The present invention relates to highly effective minimal promoters for plants, such as soybeans and rice.
Recent advances in genetic engineering have provided plant breeders and geneticists with the tools to insert or transform genes, which are selected portions of deoxyribonucleic acid (also known as DNA), into a plant in order to produce new kinds of plants known as transgenic plants. Such transgenic plants or crops can have unique characteristics or traits, including resistance to plant diseases, resistance to herbicides, resistance to insects, enhanced stability or shelf-life of the ultimate consumer product obtained from the plant and/or improvements in the nutritional value in the edible portions of the plant. Genes are made up of DNA, a complex molecule inside each plant cell that provides the instructions for all aspects of the plant""s growth. A promoter is a region on a gene where transcription factors can bind to enable the gene to xe2x80x9cexpressxe2x80x9d itself through the production of another, but smaller molecule known as messenger RNA. Messenger RNA enables the gene to xe2x80x9cdeliverxe2x80x9d its message or instructions to other parts of the plant cell in many cases by being translated into a protein. Various plant promoters have been identified and isolated from different plants, as described in various patents, such as U.S. Pat. Nos. 5,536,653; 5,589,583; 5,608,150; and 5,898,096. Although effective, such promoters have not been modified or optimized to provide enhanced or improved characteristics or traits. It would be desirable to provide plant promoters that have been modified to advantageously provide improved characteristics or traits in plants.
The present invention relates to a modified promoter that when placed upstream of a gene of interest, will cause that gene to be expressed at a high level in plant vegetative tissues. The promoter should be active during most of the plant""s developmental stages from the seedling stage to maturity.
In a first embodiment, the present invention is directed to a DNA molecule that is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9, wherein nucleotide xe2x80x9cRxe2x80x9d is xe2x80x9cAxe2x80x9d (adenine) or xe2x80x9cGxe2x80x9d guanine; and nucleotide xe2x80x9cSxe2x80x9d is xe2x80x9cCxe2x80x9d (cytosine) or xe2x80x9cGxe2x80x9d (guanine).
tatataaggaggrsttcattcccatttgaaggat [SEQ ID NO: 1]
tcctctatataaggaggrsttcattcccatttgaaggatcaata [SEQ ID NO: 2]
gtcctctatataaggaggrsttcattcccatttgaaggatcaatagttt [SEQ ID NO: 3]
ctgcagtcctctatataaggaggrsttcattcccatttgaaggatcaatagtttaaac [SEQ ID NO: 4]
More specific embodiments of SEQ ID NOS: 1-4 are shown in SEQ ID NOS: 5-8, respectively:
tatataaggaggggttcattcccatttgaaggat [SEQ ID NO: 5]
tcctctatataaggaggggttcattcccatttgaaggatcaata [SEQ ID NO: 6]
gtcctctatataaggaggggttcattcccatttgaaggatcaatagttt [SEQ ID NO: 7]
ctgcagtcctctatataaggaggggttcattcccatttgaaggatcaatagtttaaac [SEQ ID NO: 8]
aaactattgatccttcaaatgggaatgaacccctccttatatagaggactgca [SEQ ID NO: 9]
In its double stranded form, SEQ ID NOS: 1-9 are useful as minimal promoters in plants, such as dicots and monocots. For purposes of identifying the most concise minimal promoter, SEQ ID NO: 5 is preferred. For purposes of identifying the optimal minimal promoter, SEQ ID NO: 6 is preferred. For purposes of identifying the optimal minimal promoter having cloning sites for readily cloning into a vector, such as a plasmid, SEQ ID NOS: 7, 8 and 9 are preferred.
In a second embodiment, the present invention is directed toward a DNA construct comprising a DNA molecule that contains SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9. Preferably the DNA construct is a plasmid. Also preferred is that the plasmid is of the designation pPG345 or pPG346.
In a third embodiment, the present invention is directed toward a eukaryotic cell comprising a DNA molecule that contains SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9. Preferably, the eukaryotic cell is a plant cell. Also preferred is that the eukaryotic plant cell is a dicot plant cell. However, the eukaryotic plant cell may also be a monocot plant cell.
In a fourth embodiment, the present invention is directed toward a plant having eukaryotic cells comprising a DNA molecule that contains SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9. Preferably, the plant is a dicot plant, although the plant may also be a monocot plant. Also preferred is that the dicot plant is Arabidopsis thaliana. 
In a fifth embodiment, the present invention is directed toward seed capable of producing a plant having cells comprising a DNA molecule that contains SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9. Preferably, the seed is capable of producing a plant that is a dicot plant, although the seed may also be able to produce a plant that is a monocot plant.
In a sixth embodiment, the present invention is directed toward a method of controlling and/or increasing the transcription of a heterologous or homologous gene in a plant or plant tissue comprising transforming the plant or plant tissue with a DNA construct comprising a heterologous or homologous gene and a DNA molecule that is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9.